Biosynthesis of heparin. Substrate specificity of heparosan N-sulfate D-glucuronosyl 5-epimerase.

Jacobsson, Ingvar; Lindahl, Ulf; Jensen, John W.; Rodén, Lennart; Prihar, Harry S.; Feingold, David S.

doi:10.1016/s0021-9258(17)43565-4

Cited by 98 publications

(15 citation statements)

References 26 publications

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“…The uronate composition of these heparan sulphate tetrasaccharides has also been determined in the present study. From biosynthetic considerations, the internal uronate (residue 3) can only be glucuronate, as it has a GlcNAc residue on its non-reducing side (Jacobsson et al, 1984). Also, there is no evidence from the I 3C-n.m.r.…”

Section: Discussionmentioning

confidence: 99%

Very-high-field n.m.r. studies of bovine lung heparan sulphate tetrasaccharides produced by nitrous acid deaminative cleavage. Determination of saccharide sequence, uronate composition and degrees of sulphation

Sanderson¹,

Huckerby²,

Nieduszynski³

1984

Biochemical Journal

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Tetrasaccharides with the general structure UA-GlcNAc-GlcUA-aManOH (where UA represents uronate, GlcNAc N-acetylglucosamine, GlcUA glucuronate and aManOH anhydromannitol) were prepared from low-sulphated heparan sulphates of bovine lung origin by complete nitrous acid deaminative cleavage followed by reduction and fractionated by gel filtration. Ion-exchange chromatography of the tetrasaccharides yielded three major fractions in approximate yields of 37%, 45% and 14%. These were shown to be non-, mono- and di-sulphated respectively. Complete structural characterization of the tetrasaccharide fractions by quantitative high-field n.m.r. spectroscopy showed that each fraction contained only two discrete species and led to the following observations. (1) All of the uronate residues in the tetrasaccharides (and in larger oligosaccharides) are unsulphated, and hence sulphated iduronate [IdUA(2SO3)] must occur exclusively within -GlcNSO3-IdUA(2SO3)-GlcNSO3- sequences (where GlcNSO3 represents N-sulpho-glucosamine) in the parent polymers. (2) The GlcNAc residues in the tetrasaccharides are more highly C-6-O-sulphated than are the aManOH residues, and furthermore sulphation on the aManOH appears to occur only where the GlcNAc is also sulphated. (3) Where the GlcNAc is C-6-O-sulphated, iduronate is the major non-reducing terminal residue, whereas glucuronate predominates in this position if the GlcNAc is unsulphated. The quantitative data obtained are used to determine the degree of C-6-O-sulphation of glucosamine residues in specific sequences within the parent heparan sulphates.

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Section: Discussionmentioning

confidence: 99%

Very-high-field n.m.r. studies of bovine lung heparan sulphate tetrasaccharides produced by nitrous acid deaminative cleavage. Determination of saccharide sequence, uronate composition and degrees of sulphation

Sanderson¹,

Huckerby²,

Nieduszynski³

1984

Biochemical Journal

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“…2OST1 catalyses then the transfer of sulfate groups to the C2-position of both GlcA and IdoA residues, although with significantly increased efficiency towards the later, displaying around five-fold higher affinity to epimerized IdoA units ( Figure 1 ) ( 33 ). 2- O -sulfation of IdoA residues prevents their reverse epimerization into GlcA, since GLCE cannot use IdoA2S as substrate, which promotes accumulation of these residues on HS chains ( 34 ). In addition, it has been proposed that 2- O -sulfated GlcA residues (GlcA2S), generated at a smaller extent in HS chains, might result from early 2- O -sulfation, prior to the epimerization of GlcA, blocking the subsequent activity of GLCE and conversion of those residues into IdoA ( 35 ).…”

Section: Hs Biosynthesis and Modificationmentioning

confidence: 99%

“…6OST differential expression over different mouse organs was also reported, which supports tissue-dependent expression and consequent varying effects on HS structural motifs ( 40 ). 6- O -sulfation contributes to fine-tune the sequential biosynthesis of HS chains, since GlcA epimerization by GLCE is precluded by 6- O -sulfation of adjacent GlcN residues ( 34 ), and 2- O -sulfation of IdoA by 2OST1 is inhibited if the adjacent GlcN residue is 6- O -sulfated ( 36 , 41 ). These data are in line with the described stepwise mechanism underlying HS biosynthesis, further supporting that epimerization and 2- O -sulfation reactions usually take place prior to 6- O -sulfation ( Figure 1 ).…”

Section: Hs Biosynthesis and Modificationmentioning

confidence: 99%

Heparan Sulfate Biosynthesis and Sulfation Profiles as Modulators of Cancer Signalling and Progression

et al. 2021

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Heparan Sulfate Proteoglycans (HSPGs) are important cell surface and Extracellular Matrix (ECM) maestros involved in the orchestration of multiple cellular events in physiology and pathology. These glycoconjugates bind to various bioactive proteins via their Heparan Sulfate (HS) chains, but also through the protein backbone, and function as scaffolds for protein-protein interactions, modulating extracellular ligand gradients, cell signalling networks and cell-cell/cell-ECM interactions. The structural features of HS chains, including length and sulfation patterns, are crucial for the biological roles displayed by HSPGs, as these features determine HS chains binding affinities and selectivity. The large HS structural diversity results from a tightly controlled biosynthetic pathway that is differently regulated in different organs, stages of development and pathologies, including cancer. This review addresses the regulatory mechanisms underlying HS biosynthesis, with a particular focus on the catalytic activity of the enzymes responsible for HS glycan sequences and sulfation motifs, namely D-Glucuronyl C5-Epimerase, N- and O-Sulfotransferases. Moreover, we provide insights on the impact of different HS structural epitopes over HSPG-protein interactions and cell signalling, as well as on the effects of deregulated expression of HS modifying enzymes in the development and progression of cancer. Finally, we discuss the clinical potential of HS biosynthetic enzymes as novel targets for therapy, and highlight the importance of developing new HS-based tools for better patients’ stratification and cancer treatment.

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“…the heparin proteoglycan, through a series of modification reactions that include deacetylation of N-acetylglucosamine residues, sulphation of the resulting free amino groups, C-5-epimerization of D-glucuronic acid to L-iduronic acid units and finally O-sulphation at various positions (Kusche et al, 1988; for reviews see also Lindahl et al, 1986;Lindahl & Kjellen, 1987;Lindahl, 1989). The reactions occur rapidly, the entire series of modifications being completed within 1 min for an individual polysaccharide chain (H66k et al, 1975), yet proceed in a stepwise manner, such that certain structural modifications are introduced before the subsequent reactions can be initiated (Jacobs-son & Riesenfeld et al, 1980Riesenfeld et al, , 1982Jacobsson et al, 1979Jacobsson et al, , 1984.…”

Section: Introductionmentioning

confidence: 99%

Biosynthesis of heparin. Relationship between the polymerization and sulphation processes

Lidholt

Kjellén

Lindahl

1989

Biochemical Journal

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Incubation of a mouse mastocytoma microsomal fraction with UDP-[3H]GlcA and UDP-GlcNAc yielded proteoglycans containing non-sulphated polysaccharide chains. Similar incubations performed in the presence of sulphate donor 3'-phosphoadenosine 5'-phosphosulphate (PAPS) produced both sulphated and non-sulphated proteoglycans, which were separated by chromatography on DEAE-cellulose Analysis by gel chromatography of single polysaccharide chains, released from the proteoglycans by alkali treatment, showed that the non-sulphated chains produced during incubation for 5 min or 25 min, either in the absence or in the presence of PAPS, were of fairly small molecular size, with an average peak Mr of approx. 10 x 10(3)-15 x 10(3). In contrast, the sulphated chains exceeded Mr 100 x 10(3) Pulse-chase experiments suggested that sulphated chains were capable of further elongation. These results indicate that sulphation promotes, by so far unknown mechanisms, further chain elongation. Sulphated proteoglycan (retarded on DEAE-cellulose chromatography) isolated after similar incubation of the microsomal fraction for 1 min only was found to contain a mixture of sulphated and virtually non-sulphated polysaccharide chains. However, when [35S]PAPS was included in the incubations, some 35S was found to be associated, essentially as N-sulphate groups, also with the latter type of chains, preferentially the high-Mr fraction. These results are interpreted in terms of a biosynthetic model by which the heparin proteoglycan is generated through transient interactions of macromolecular intermediates with distinctly separate complexes of membranebound enzymes.

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Biosynthesis of heparin. Substrate specificity of heparosan N-sulfate D-glucuronosyl 5-epimerase.

Cited by 98 publications

References 26 publications

Very-high-field n.m.r. studies of bovine lung heparan sulphate tetrasaccharides produced by nitrous acid deaminative cleavage. Determination of saccharide sequence, uronate composition and degrees of sulphation

Very-high-field n.m.r. studies of bovine lung heparan sulphate tetrasaccharides produced by nitrous acid deaminative cleavage. Determination of saccharide sequence, uronate composition and degrees of sulphation

Heparan Sulfate Biosynthesis and Sulfation Profiles as Modulators of Cancer Signalling and Progression

Biosynthesis of heparin. Relationship between the polymerization and sulphation processes

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